The present invention relates to the production of the seedlings or plantlets of Eleutherococcus senticosus(Siberian ginseng). More particularly, the present invention relates to a method for mass producing the seedlings or plantlets of Eleutherococcus senticosus through the somatic embryogenesis of Eleutherococcus senticosus/embryogenic cells by using bioreactors. In one aspect, the seedlings or plantlets of Eleutherococcus senticosus are used as health foods or health aid foods.
Eleutherococcus senticosus, which is known as an efficacious medicinal herb all over the world, grows naturally in Korea, China, Japan, and Siberia.
Since the first report of disclosing the value of naturally occurring Eleutherococcus senticosus (alias Acanthopanax senticosus) as a medicine, researchers in many countries, including Russia, have made efforts to reveal its medicinal components. As a result, acanthoic acid, which is evaluated as being five times more potent in anti-inflammatory activity than is aspirin, xcex2-sitosterol, eleutheroside A-G, and stigmasterol were extracted from Eleutherococcus senticosus. By virtue of the cooperation of these medicinal components, Eleutherococcus senticosus is now found to effect invigoration, life extension and homeostasis in addition to showing therapeutic activity against hypertension, diabetes, cancer, inflammation, fever and pain, neuralgia, etc, as do Koran ginsengs. Particularly, Eleutherococcus senticosus is known to be of sovereign remedy in relieving physical and mental fatigue.
A research report discloses that eleutheroside E, which shows various physiologically active effects, is contained at an amount 1.7xcx9c5.5 times greater in the Eleuthrococcus senticosus native to Korea than in other native Acanthopanax spp. such as Acanthopanax chiisanesis, Acanthopanax seoulense, Acanthopanax sieboldianum, and Acanthopanax koreanum (Park, H. K., a thesis for a doctorate, Chonbuk National University, 1997, Korea).
However, Eleutherococcus senticosus is difficult for general farm households to seed because of its stringent weather conditions. In addition, the cutting of the plant is not effective for proliferation. Also, because medicinal materials are taken from the velamen and the bark of Eleutherococcus senticosus, it takes a long period of time for this shrub to be cultivated to useful extent. What is worse, once the velamen and the bark are taken off, the plant no more maintains its existence. That is, this plant cannot afford reproductive provisions of the medicinal materials, but gives only one chance for getting the medicinal materials.
In the case of ginsengs, success was brought about in mass culturing their cells in culture vessels, enabling the extraction of saponins therefrom (Nitto Denko, Japan). In addition, the cultured bodies are commercially sold as a health food after being dried and powdered in their entirety without processing. As for yew, its cells are cultured to obtain paclitaxel, an anticancer compound.
However, there have been found almost no cases in which the seedlings or plantlets regenerated though cell culturing are mass produced for commercialization as health foods, as in the present invention to be later described in detail.
There are reports regarding the generation of adventitious buds through tissue culture or the regeneration of plants through somatic embryogenesis (Plant Cell Reports 9: 514-516, 1991; Korea J. Plant Tissue Culture 20:216-226, 1993). Achieved only in small scales on agar medium, the techniques disclosed in the reports are different from the present invention in which embryogenic cell strains are established in a broth and used to produce seedlings or plantlets in a large quantity. Until the present invention, there has been reported no research on the system for commercializing Eleutherococcus senticosus as health foods or health aid foods by mass producing Eleutherococcus senticosus through tissue culture.
The present invention provides methods that produce seedlings or plantlets of Eleutherococcus senticosus in a highly efficient manner. Moreover, the seedlings or plantlets can be used as health foods.
Eleutherococcus senticosus/embryogenic cells (Accession number given by the INTERNATIONAL DEPOSITARY AUTHORITY: KCTC 0504BP) are suspension-cultured in MS (Murashige and Skoog) liquid medium under a dark condition to allow the mass proliferation of the embryogenic cells. The invention provides a method for culturing Eleutherococcus senticosus seedlings or plantlets, in which embryogenic cells are subcultured to homogeneous sizes of somatic embryos in liquid medium and the somatic embryos in a bioreactor equipped with an airlift are cultured to produce the seedlings or plantlets. The subculturing is carried out repeatedly every 2xcx9c3 weeks.
In one embodiment, the proliferated cells are classified according to sizes by filtering them through a network ranging, in pore size, from 150 to 300 xcexcm before the classification. In another embodiment, the embryogenic cells in the filtrate are further suspension-cultured in liquid medium while the embryogenic cell clusters entrapped in the network are subjected to culturing in the bioreactors of liquid medium to be regenerated to the seedlings or plantlets.